• BMB Reports
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• v.30 no.4
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• pp.246-252
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• 1997

Malonate decarboxylase from Acinetobacter calcoaceticus is shown to have malonyl-CoA: acetate CoA transferase. acetyl-CoA: malonate CoA transferase, and malonyl-CoA decarboxylase activity. These enzyme activities were elucidated by isotope exchange reactions. The enzyme modified by N-ethylmaleimide completely lost its malonate decarboxylase activity, whereas it still kept CoA transferases and malonyl-CoA decarboxylase activities. The existence of CoA transferases and malonyl-CoA decarboxylase activity is clear, but their physiological significance is obscure. The catalytic reactions for two eoA transfers and malonyl-CoA decarboxylation proceed via a cyclic mechanism, which is through two covalent intermediates, enzyme-Smalonyl and enzyme-S-acetyL proposed for malonate decarboxylation of the enzyme.

• BMB Reports
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• v.32 no.4
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• pp.414-418
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• 1999

A novel gene for malonyl-CoA decarboxylase was discovered in the mat operon, which encodes a set of genes involved in the malonate metabolism of Rhizobium trifolii (An and Kim, 1998). The subunit mass determined by SDS-PAGE was 53 kDa, which correspond to the deduced mass from the sequence data. The molecular mass of the native enzyme determined by field flow fractionation was 208 kDa, indicating that R. trifolii malonyl-CoA decarboxylase is homotetrameric. R. trifolii malonyl-CoA decarboxylase converted malonyl-CoA to acetyl-CoA with a specific activity of 100 unit/mg protein. Methylmalonyl-CoA was decarboxylated with a specific activity of 0.1 unit/mg protein. p-Chloromercuribenzoate inhibited this enzyme activity, suggesting that thiol group(s) is(are) essential for this enzyme catalysis. Database analysis showed that malonyl-CoA decarboxylase from R. trifolii shared 32.7% and 28.1% identity in amino acid sequence with those from goose and human, respectively, and it would be located in the cytoplasm. However, there is no sequence homology between this enzyme and that from Saccharopolyspora erythreus, suggesting that malonyl-CoA decarboxylases from human, goose, and R. trifolii are in the same class, whereas that from S. erythreus is in a different class or even a different enzyme, methylmalonyl-CoA decarboxylase. According to the homology analysis, Cys-214 among three cysteine residues in the enzyme was found in the homologous region, suggesting that the cysteine was located at or near the active site and plays a critical role in catalysis.

• BMB Reports
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• v.35 no.2
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• pp.213-219
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• 2002

Malonyl-CoA decarboxylase (E.C.4.1.1.9) catalyzes the conversion of malonyl-CoA to acetyl-CoA. Although the metabolic role of this enzyme has not been fully defined, it has been reported that its deficiency is associated with mild mental retardation, seizures, hypotonia, cadiomyopathy, developmental delay, vomiting, hypoglycemia, metabolic acidosis, and malonic aciduria. Here, we isolated a cDNA clone for malonyl CoA decarboxylase from a rat brain cDNA library, expressed it in E. coli, and characterized its biochemical properties. The full-length cDNA contained a single open-reading frame that encoded 491 amino acid residues with a calculated molecular weight of 54, 762 Da. Its deduced amino acid sequence revealed a 65.6% identity to that from the goose uropigial gland. The sequence of the first 38 amino acids represents a putative mitochondrial targeting sequence, and the last 3 amino acid sequences (SKL) represent peroxisomal targeting ones. The expression of malonyl CoA decarboxylase was observed over a wide range of tissues as a single transcript of 2.0 kb in size. The recombinant protein that was expressed in E. coli was used to characterize the biochemical properties, which showed a typical Michaelis-Menten substrate saturation pattern. The $K_m$ and $V_{max}$ were calculated to be $68\;{\mu}M$ and $42.6\;{\mu}mol/min/mg$, respectively.

• Bulletin of the Korean Chemical Society
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• v.20 no.10
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• pp.1159-1164
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• 1999

The study investigated the effect of carrier composition (ionic strength and pH) on the retention of various proteins in flow field-flow fractionation (Flow FFF) as well as the conformational change of Bovine Serum Albumin (BSA) with urea concentration, storage time and temperature. The study found that the retention of protein in Flow FFF increased with the ionic strength of the carrier liquid. Most proteins were well solubilized at pH = 7-8. The hydrodynamic diameters obtained from Flow FFF retention data agree well with theoretical values. The retention increased and the peak shape became distorted at extreme pH conditions of the carrier solution. The selected carrier composition for comparison between the literature value of proteins was 0.05 M tris buffer solution with a pH of 8. Storing BSA at 4 ±2℃ over a period of three months resulted in slow dimerization. Also, in case of the storage of BSA at 37 ±5℃ for one week, the retention of both BSA monomer and dimer increased with the urea concentration. Finally, the structural composition of specific enzymes: malonyl-CoA decarboxylase (MCDC) and malonyl-CoA synthesis (MCS) was determined by using Flow FFF at specific carrier solutions. The molecular weight of the natural MCDC was determined to be 208 kDa, which means it is a homotetramer, while that of the MCS was determined to be 47 kDa, which means it is a monomer.

• BMB Reports
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• v.35 no.5
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• pp.443-451
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• 2002

Malonate is a three-carbon dicarboxylic acid. It is well known as a competitive inhibitor of succinate dehydrogenase. It occurs naturally in biological systems, such as legumes and developing rat brains, which indicates that it may play an important role in symbiotic nitrogen metabolism and brain development. Recently, enzymes that are related to malonate metabolism were discovered and characterized. The genes that encode the enzymes were isolated, and the regulation of their expression was also studied. The mutant bacteria, in which the malonate-metabolizing gene was deleted, lost its primary function, symbiosis, between Rhizobium leguminosarium bv trifolii and clover. This suggests that malonate metabolism is essential in symbiotic nitrogen metabolism, at least in clover nodules. In addition to these, the genes matB and matC have been successfully used for generation of the industrial strain of Streptomyces for the production of antibiotics.